PULLULAN as Cosmetic Ingredient. Hayashibara Biochemical Laboratories, Inc.

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1 PULLULAN as Cosmetic Ingredient Hayashibara Biochemical Laboratories, Inc.

2 What is Pullulan? Chemical Structure Maltotriose Natural polysaccharide derived from starch Manufactured by fermentation process An linear polysaccharide consisting of repeating maltotriose units linked by α-1,6-glycosidic bonds.

3 Properties

4 Contents 1. Solubility 2. Lubricity 3. Film forming property 4. Adhesiveness 5. Low assimilating property 6. Non- Cariogenicity

5 Property 1. Solubility Soluble in both cold water and hot water Insoluble in ethanol but soluble in polyhydric alcohol, propylene glycol and 1,3-butylene glycol

6 Property 2. Lubricity Pullulan has lubricity unless a polymer Example Lotion Cream Effect Improvement for Smooth and Soft Texture

7 Property 3. Film forming property Pullulan has excellent water-soluble film forming property after drying the aqueous solution. This film is transparent. Example Face mask Hair styling Effect Rip hard Protect hair style

8 Property 4. Adhesiveness High Adhesiveness Pullulan is extremely adhesive, even by a spray of solution, it glues to other materials. Available for tablets or binder of granule

9 Property 5. Assimilating property with resident microbic on skin Aerobic condition Anaerobic condition Aerobic condition Anaerobic condition Brevibacterium epidermidis Corynebacterium xerosis Micrococcus kristinae Staphylococcus aureus Staphylococcus epidermidis Lactobacillus pentosus Propionibacterium acnes Residue of each saccharide (%) Pullulan Glucose Bacteria were each cultured in medium containing Pullulan or glucose each under Aerobic and Anaerobic conditions. The assimilating property was determined by measuring the residue of each saccharide.

10 Property 6. Cariogenicity 1 Acid fermentation test Streptococcus sobrinus Streptococcus mutans ph Critical Critical ph ph of of enamel decalcification Critical ph of enamel decalcification Control 5.5 Sucrose Pullulan 5.0 ph Reaction time (min) Reaction time (min) S. Sobrinus and S. mutans were cultured in 0.02M saccharide containing media and kept at 37 C The ph of each media as acid production was measured over time.

11 Property 6. Cariogenicity 2 Insoluble glucan Streptococcus sobrinus Streptococcus mutans Sucrose スクロース Sucrose スクロース Pullulan プルラン Pullulan プルラン Isomaltose イソマルトース Isomaltose イソマルトース Insoluble 不溶性グルカン glucan (mg) Insoluble 不溶性グルカン glucan (mg) The glucosyltransferase delived from S. sobrinus and S. mutans and saccharides were respectively cultured in phosphate buffer solution at 37 C for 16 hours and the insoluble glucan was measured.

12 Application to Cosmetics

13 for Skin-care Cosmetics: Extensibility 1 Smoothness on Skin applied Pullulan containing Cream Extensibity 1.40 good Mean coefficient of friction (MIU) Averaged coefficient of friction (MIU) ** Control Pullulan Carbomer A Carbomer B CarbomerC 1. Put a few drops of cream (0.4ml) containing polymer (0.3%) on the artificial leather. 2. Measure the load by making contact shoe slide in the same direction. 3. Repeat 13 times and calculate the average of the value between 10th and 13th 1). 4. The smoothness was confirmed by Averaged friction coefficient (MIU). **:p < 0.01 vs. Control, Carbomer A, B and C (Scheffe's post hoc test) 1) K. Suganuma, Fragrance Journal., 2, (1995)

14 for Skin-care Cosmetics: Availability 2 Smoothness on Skin applied Pullulan containing Cream Variance 摩擦係数変動 of of friction friction coefficient coefficient (MMD) (MMD) Firm Oily Oily Hydrate Dry Mean 平均摩擦係数 coefficient of (MIU) friction Control Pullulan Carbomer A Carbomer B Carbomer C 1. Put 0.4ml of cream containing polymer (0.3%) on the artificial leather. 2. Measure the load 2 hours after application. 3. The smoothness was confirmed by Averaged friction coefficient (MIU) and Variance of friction coefficient (MMD) 2). 2) Egawa M., J. Soc. Cosmet. Chem. Japan, 37, (2003)

15 for Skin-care Cosmetics: Face Musk Smoothness on Skin applied Pullulan containing Cream Available for both Transparent-type and Foam-type Water solubility Oily substances are also available in the Musk Preservatives free Apply the Musk on face after moistening with face lotion or essence.

16 Overview

17 Availability for Cosmetics Lubricity Lotion Cream Shampoo Humid Condition Lotion Cream Musk Film forming Property Dry Condition Adhesiveness Powder

18 Regulatory Affair Ingredient name PULLULAN Labeling name PULLULAN INCI name PULLULAN Pullulan is also available for additive for Quasi-Drug in Japan.

19 Specifications Description Specifications White to light yellowishwhite, odorless or slight specific odor powder Method and Description The Japanese Standards of Quasi- Drug Ingredients (2006) Identification (1) Give viscous solution Condition of the solution Identification (2) Lose its viscosity React with Pullulanase Identification (3) Cause white precipitation React with PEG 600 ph JSQI GTP (1) Heavy Metal Not more than 20ppm JSQI GTP, 2nd method (2) Arsenic Not more than 2ppm JSQI GTP, 3rd method, apparatus B (3) Protein Not more than 0.3 % JSQI GTP, Nitrogen determination, 1st method Loss on drying Not more than 8.0 % JSQI GTP (1g, under reduced pressure, 90 C, 6h) Residue on ignition Not more than 5.0 % JSQI GTP (1stmethod,2g)

20 Safety Species Route & Duration Dosage Reasult Acute toxicity Mouse Pass oral, Single g/kg LD 50 >14280 Sub-Acute toxicity Rat Pass oral, 13 weeks 7.9 g/kg/day NAD Chronic Toxicity Rat Pass oral, 62 weeks g/kg/day NAD Sensitization Mouse & Guinea pig Passive cutaneous & Active anaphylaxis test Approx. 150 μg/kg Negative Once a week, 3 weeks Ocular irritancy test bovine cornea 4 hours 20 W/V % Negative Chorioallantois of fertilized chicken egg 5 minutes 0.3 g, Powder Negative Mutagenicity test murine Bacillus typhosus 10 mg/ plate Negative Patch test Human Transdermal, 24 hours 3 times a week, 3 weeks 25% aq. solution 0.2 ml NAD

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